Background & Objectives: Epstein-Barr virus (EBV) is a tumorigenic herpes virus and belong to DNA viruses that infects and persists in B lymphocytes in the majority of humans and also is associated with a portion of cases of Hodgkin lymphoma (HL). The aim of the present study was to investigate the presence of Epstein-Barr virus in Hodgkin's lymphoma through EBERs in situ hybridization (EBERs-ISH) in children Iranian. Materials & Methods: In this study, 16 Hodgkin's lymphoma tissue samples formalin-fixed paraffin embedded blocks collected from the archive of departments country hospitals. Samples were obtained from the tissue cancer biopsies, The presence of RNA transcripts encoded by Epstein-Barr virus were analyzed with EBERs in situ hybridization (EBERs-ISH). Data analyzed by SPSS16 statistical software, Fisher's exact test. Results: The presence of Epstein-Barr virus (EBV) in 16 formalin-fixed, paraffin-embedded samples of Hodgkin's disease (HD) was investigated by EBERs in situ hybridization. 12 samples were from male (75%) and 4 samples were from female (25%). Age range of Patients was 4 -12 years (with a median of 8 years). In 12 specimens (75%) Epstein-Barr virus was present. The positive cases included 9 male samples (75%) and 3 female samples (75%). Fisher's exact test showed statistically no significant difference between sex (p-value: 1.000). Epstein-Barr virus was present in different subtypes of Hodgkin's lymphoma. 80% (4 of 5) of Mixed Cellularity (MC), 67% (6 of 9) of Nodular Sclerosis (NS) and 100% (2 of 2) of Lymphocyte Predominance (LP) subtypes were positive. There is not significant different between subtypes (p-value: 0.826). The presence of Epstein-Barr virus in the age groups of 4-7 and 8-12 years were 71.5% (5 of 7) and 77.8% (7 of 9), respectively. There is not significant different between age groups (p-value: 1.000). Conclusion: Study results show strong association between Epstein-Barr virus and Hodgkin lymphoma in Iranian children. Epstein-Barr virus and Hodgkin lymphoma association in Iranian children show similar pattern with other developing countries.